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Neuroscience JoVE (Open Access) Citable · DOI

Axon Stretch Growth: The Mechanotransduction of Neuronal Growth

DOI: 10.3791/2753-v
What you'll learn
  • ✓ Set up and operate an axon stretch growth bioreactor system
  • ✓ Culture primary neurons and initiate mechanical stretch protocols
  • ✓ Monitor axon elongation using time-lapse microscopy and mechanotransduction principles
Protocol

A unique tissue engineering method was developed to elongate numerous nerve fibers in culture by recapitulating axon stretch growth; a form of nervous system growth whereby nerves elongate in conjunction with growth of the enlarging body.

Difficulty
advanced
Total time
~5-7 days (culture preparation + stretch protocol + imaging)
Model organism
Primary neuronal culture (origin species not specified in abstract)
Biosafety
BSL-1

Steps

1
Overview of axon stretch growth bioreactor system

Learn the bioreactor design and principles for recapitulating nervous system growth through mechanical stretch. Understand how the system mimics physiological axon elongation during body development.

▶ 01:41
2
Prepare bioreactor chamber for neuronal culture

Assemble and prepare the bioreactor chamber components according to tissue engineering specifications. Ensure sterility and proper configuration for subsequent neuronal seeding.

▶ 03:06
3
Culture primary neurons in bioreactor

Seed and maintain primary neuronal cultures within the prepared bioreactor chamber. Establish baseline culture conditions before mechanical stretch application.

▶ 05:49
4
Apply axon stretch growth mechanical protocol

Execute the stretch protocol to elongate axons in culture. Monitor mechanotransduction response during the stretching phase.

▶ 07:20
5
Acquire time-lapse imaging of axon elongation

Set up and perform time-lapse microscopy to visualize axon growth dynamics during mechanical stretch. Document morphological changes over the experimental period.

▶ 08:48
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